Detects rat ICAM-1/CD54 in direct ELISAs and Western blots. In direct ELISAs, approximately 40% cross-reactivity with recombinant mouse (rm) ICAM-1 and 5% cross-reactivity with recombinant human (rh) ICAM-1, rhICAM‑3, and rmICAM‑2 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Rat ICAM-1/CD54 Antibody

Detection of Rat ICAM‑1/CD54 by Western Blot.

Western blot shows lysates of rat liver tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Rat ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF583) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for ICAM-1/CD54 at approximately 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

ICAM-1/CD54 in Rat Brain.

ICAM-1/CD54 was detected in perfusion fixed frozen sections of rat brain using Goat Anti-Rat ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF583) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Rat ICAM-1 / CD54 ELISA Standard Curve

Recombinant Rat ICAM-1/CD54 Fc Chimera (Catalog # 583-IC) was serially diluted and captured by Mouse Anti-Rat ICAM-1/CD54 Monoclonal Antibody (Catalog # MAB5832) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat ICAM-1/CD54 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF583) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

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Applications for Rat ICAM-1/CD54 Antibody

Application

Recommended Usage

Adhesion Blockade

The adhesion of HSB2 human peripheral blood acute lymphoblastic leukemia cells (5 x 10⁴ cells/well) to immobilized Recombinant Rat ICAM-1 Fc Chimera (Catalog # 583-IC, 12.5 µg/mL, 100 µL/well) was maximally inhibited (80-100%) by 25 µg/mL of the antibody.

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of rat brain (vasculature)

Western Blot

0.2 µg/mL
Sample: Rat liver tissue

Reviewed Applications

Read 1 review rated 5 using AF583 in the following applications:

Immunohistochemistry (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: ICAM-1/CD54

Intercellular Adhesion Molecule-1 (ICAM-1, CD54), binds the leukocyte integrins LFA-1 and Mac-1. ICAM-1 expression is weak on leukocytes, epithelial and resting endothelial cells, as well as some other cell types, but expression can be stimulated by IFN-gamma, TNF-alpha, IL-1 beta, and lipolysaccharide (LPS). Rat and human ICAM-1 share approximately 52% amino acid identity. Rat and mouse ICAM-1 share approximately 52% amino acid identity.

Soluble ICAM-1 is found in a biologically active form in serum, probably as a result of proteolytic cleavage from the cell surface, and is elevated in patients with various inflammatory syndromes such as septic shock, leukocyte adhesion deficiency syndrome (LAD), cancer, and transplantation.

References

Pigott, R. and C. Power (1993) in The Adhesion Molecule Facts Book, p. 74. Academic Press.
Siu, G. et al. (1989) J. Immunol. 143:3813.
Ballantyne, C.M. et al. (1989) Nuc. Acid. Res. 17:5853.

Long Name

Intercellular Adhesion Molecule 1

Alternate Names

CD54, ICAM1

Entrez Gene IDs

3383 (Human); 15894 (Mouse); 25464 (Rat)

Gene Symbol

ICAM1

UniProt

Q00238

Additional ICAM-1/CD54 Products

Product Documents for Rat ICAM-1/CD54 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Rat ICAM-1/CD54 Antibody

For research use only

Citations
Reviews

Rat ICAM-1/CD54 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF583

Key Product Details

Species Reactivity

Validated:

Cited:

Applications